Optimizing α-amylase production by an extremely halophilic archaeon Haloferax mucosum MS1.4, using response surface methodology

Neethu, K P and Sobhana, K S and Keerthi, K R and Varghese, Eldho and Ranjith, L and Jasmine, S and Joshi, K K and George, Grinson (2025) Optimizing α-amylase production by an extremely halophilic archaeon Haloferax mucosum MS1.4, using response surface methodology. Journal of Microbiological Methods. pp. 1-10. ISSN 1872-8359

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Abstract

This study investigated α-amylase production potential of the haloarchaeal strain Haloferax mucosum MS1.4 (NCBI GenBank Acc. No. OL780830), isolated from a solar saltpan located in Tamil Nadu, southern India. The enzyme was partially purified and characterized, employing SDS-PAGE analysis indicating a molecular mass of approximately 67 kDa. In order to optimize the conditions for maximizing α-amylase production, a two-step statistical approach was adopted. The initial phase involved using a Plackett-Burman Design (PBD) to determine the primary factors affecting enzyme production. PBD identified incubation time, starch concentration and NaCl concentration as the most significant variables influencing enzyme yield. These factors were further optimized using Box-Behnken Design (BBD) under Response Surface Methodology (RSM). Maximum α-amylase production of 955.97 units min−1 was obtained under the optimized conditions of 1.62 % starch; 29.88 % NaCl and an incubation period of 144 h (6 days). The findings highlight the potential of H. mucosum MS1.4 as a promising candidate for industrial enzyme production under high-saline conditions.

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